Identification of individual prokaryotic cells by using enzyme-labelled, rRNA-targeted oligonucleotide probes

Article Abstract:

Oligonucleotide probes specific to complementary rRNA sequences of the domains Bacteria and Archaea were fused to horseradish peroxidase (HRP) in an attempt to evaluate the use of HRP-conjugated probes for the identification of single bacterial cells using light microscopy. The procedure also involved the use of lysozyme-EDTA to permeabilize gram-negative cells, although this was incorporated in the hybridization step for some archaebacterial cells. The procedure was effective for gram-negative and archaebacterial cells. However, the gram-positive cells are resistant to the penetration of the HRP-conjugated probe.

Gram-negative bacteria, Identification and classification, Archaeabacteria, Microscope and microscopy, Microscopes, Microscopy, Archaea

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Dual staining of natural bacterioplankton with 4',6-diamino-2-phenylindole and fluorescent oligonucleotide probes targeting kingdom-level 16S rRNA sequences

Article Abstract:

A dual staining technique using 4', 6-diamidino-2-phenylindole and fluoresent oligonucleotide probes specific for kingdom-level 16S rRNA sequence was developed. The method increased the sensitivity and visibility of natural bacterioplankton, thus allowing the microscopic characterization and quantification of specific phylogenetic group in dilute aquatic communities. The specific oligonucleotide probes could also be used at other taxonomic levels.

Innovations, Stains and staining (Microscopy), Stains (Microscopy), Plankton research

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Monitoring the enrichment and isolation of sulfate reducing bacteria by using oligonucleotide hybridization probes designed from environmentally derived 16S rRNA sequences

Article Abstract:

The enrichment and isolation of multispecies anaerobic reactor-derived sulfate reducing bacterium by fluorescent oligonucleotide hybridization probes are discussed. Growth studies, cell fixation, hybridization photomicroscopy, 16s rRna sequencing and phlogenetic analysis were used on a molecular isolate related to Desulfovibrio vulgaris. The utility of complementing molecular to classical microbiology was emphasized.

Research, Bacteria, RNA, Hybridization

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